Figure 1.

Generation and validation of gonadotrope-specific Smad2/3-knockout mice. A, B) YFP⁺ and YFP⁻ cells were isolated from GRIC-YFP and S2/3cKO-YFP mice by FACS. Genomic DNA was extracted and analyzed by genotyping PCR for the wild-type (WT), floxed (flox), and recombined (rec.) alleles of Smad2 (A) and Smad3 (B). Data are representative from 1 of 3 sorting experiments. C, D) cDNA was prepared from total RNA from sorted cells and analyzed by qPCR using primers overlapping the deleted exons in Smad2 (C: forward primer in exon 10; reverse primers in exon 11) and Smad3 (D: forward primer in exon 3; reverse primer in exon 4). Smad2 and Smad3 transcript levels were normalized to the levels of the housekeeping gene Rpl19. Data represent means ± se from 3 independent sorting experiments assayed in triplicate. Bars with different symbols differ significantly.